This research aimed to evaluate the antibacterial effectiveness of a herbal hand sanitizer from a combination of methanol extracts of Asian Sweet flag (Acorus calamus L.) and fragrant pandan (Pandanus amaryllifolius Roxb. ex Lindl.) leaves, and to identify the bioactive compounds by GC-MS analysis. Asian sweet flag (Dlingo in Javanese) and pandan leaves were macerated in methanol. The extracts were subjected to phytochemical screening and formulated into hand sanitizer preparations with concentrations of 25%, 50%, 75%, and 100%, including a single 100% formulation of each extract. The disc diffusion method was used to investigate antibacterial activity against Staphylococcus aureus and Escherichia coli, while bioactive compounds were identified by GC-MS. The results showed that the extraction yielded 17.75% for dlingo leaves and 18% for pandan leaves. Phytochemical analysis identified flavonoids, tannins, alkaloids, and saponins in both extracts. All formulations inhibited S. aureus and E. coli, with the highest activity in the 100% combination formulation, producing inhibition zones of 9.60 mm and 6.47 mm. GC-MS analysis identified 196 compounds dominated by phenolics, aromatic aldehydes, benzofurans, and long-chain fatty acids with antibacterial potential. The study concludes that the combination of dlingo and pandan leaf extracts has potential as a natural active ingredient in herbal hand sanitizer formulations.
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